Scientific Program

Conference Series Ltd invites all the participants across the globe to attend 4th World Congress on Virology San Antonio, USA.

Day 1 :

  • Track 2: Viral Breakout: Prevention and Measures
Location: San Antonio, USA

Session Introduction

Nguyen Trong Binh

Biotechnology center of Ho Chi Minh City, Vietnam

Title: Genetic analyses of the function of PB1 subunit of the influenza virus RNA-dependent RNA polymerase

Time : 13:05-13:25

Biography:

Nguyen Trong Binh completed PhD, March, 2014 from University of Tsukuba, Japan. At present, I'm a group leader of vaccine development against viruses, Biotechnology center of Ho Chi Minh City, Vietnam.

Abstract:

The influenza virus genome forms viral ribonucleoprotein (vRNP) complexes with nucleoprotein and viral RNA polymerases, PB1, PB2, and PA subunits. The vRNP complex catalyzes both replication and transcription reactions. PB1 functions as a catalytic subunit of RNA polymerase and contains the highly conserved motifs of RNA-dependent RNA polymerases. The C-terminal region of PB1 between amino acid (a.a.) positions 494-757 contains a putative vRNA promoter binding site, while the N-terminal region of PB1 between a.a. positions 1-83 contains both putative vRNA and cRNA promoter binding sites and a PA binding site. However, except for the PA binding site, the crystal structure and the function of the N-terminal region of PB1 are poorly understood. Here, we have examined the functional structure of the N-terminal region of PB1. The regions between a.a. positions 1-50 are highly conserved between influenza A and B viruses, but amino acids at positions 16, 27, and 44 are different between two viruses. To elucidate the functional importance of these amino acids in replication and transcription of the viral genome, we generated viruses containing mutations at these positions by reverse genetics and examined the replication and transcription activities of these mutants. We found that a.a. positions 27 and 44 are responsible for the viral replication activity but not transcription activity. Furthermore, we analyzed the function of PB1 subunit based on nucleotide recognition using ribavirin. We found that asparagine at position 27 in PB1 is responsible for ribavirin resistance.

Ayae Honda

Hosei University, Japan

Title: Influenza virus infection rise the temperature of cell

Time : 12:45-13:05

Biography:

Ayae Honda is working as a researcher in Hosei University, Japan.

Abstract:

The genome of Influenza virus is negative strandedRNA and about 14000 nucleotides. The replication takes place in nucleus using own RNA dependent RNA polymerase.We prepared to temperature sensor to examine the cell temperature. Using this temperature sensor we measured influenza virus infected- and uninfected- cells. Result showed thatthe temperature of influenza virus infection rosecell temperatureto about 5oC. To understand this phenomenon we assayed ATP level in the cell. ATP content of the virus infected cells rose up to 2hpi, and then decreased. RT-PCR for mRNA showed that the mRNA was increased 1000 fold up to 4hpi. We also assayed the viral protein content at each time after virus infection. At 6-8 hpi, most of viral proteins were detected by LC/MS correlated to the result of RT-PCR. This result indicated that a large amount of ATP consumption caused the rise of temperature inner cell.

  • Trackl 3: Human Viral Diseases Affecting Afro-Asian Continents
Location: San Antonio, USA

Session Introduction

Louise Cosby

Queen’s University Belfast
United Kingdom.

Title: Measles virus: Complications of infection and future threats from related veterinary viruses

Time : 14:25-14:45

Speaker
Biography:

S. Louise Cosby is a graduate (B.Sc. and Ph.D. in Microbiology) of Queen’s University Belfast and a member of staff for over 20 year. She is a Fellow of the Royal College of Pathologists (London) and a Fellow of the Society of Biology (UK). She was a visiting associate professor in Cornell University USA and a visiting scientist at the institute of Animal Health, Pirbright, UK. She was appointed to the Chair of Microbiology in 2002.); Previous/present board membership of the Biochemistry and Cell Biology Committee, BBSRC, UK; Biochemistry Board, Science Foundation Ireland (previous chair of a Bioscience panel); Infections and Immunity and Host Defence Panel, Health Research Board, Ireland; GersonLerman’s Group’s Healthcare Advisor’s Board, USA. SLC is an Associate Editor for the Journal of Neurovirology;a review editor for Frontiers in Microbiology; an external assessor for appointments and promotions in Medical Microbiology, University of Malaysia; assessor for grant applications for the European Commission.

Abstract:

The World Health Organisation has set regional elimination goals for measles virus (MV) eradication to be achieved by 2020 or earlier. A major question is whether an opportunity for veterinary virus infection of humans may arise when MV is eradicated and if vaccination is discontinued. Lessons have been learned from animal to human virus transmission i.e. human immunodeficiency virus (HIV) and more recently from severe acute respiratory syndrome (SARS) and avian influenza virus infections. We are therefore alerted to the risk of zoonosis from the closely related veterinary viruses in the same morbillivirus genus as MV. Of most concern with regard to zoonosis is the recently reported fatal infection of primates with canine distemper virus (CDV). These animals displayed neurological symptoms and pathology similar to CDV infection in dogs.To allow a morbillivirus to initially infectanother species, specific cell entry receptors must be present in relevant cell types and tissues of the host. Signalling lymphocyte activation molecule (SLAM) found on immune cell types has been identified as a receptor for all morbillviruses. More recently poliovirus receptor related 4 (PVRL4, also known as nectin 4), found on the basolateral surface of polarised epithelial cells, has been identified as a receptor for MV and for CDV in dogs. We have recently reported that PVRL4 is up-regulated in human brain endothelial cells in culture following MV infection(Abdullah et al. 2013, J. NeuropatholExpNeurol 72: 681-696).and therefore could have a role in cell entry into the CNS through the blood brain barrier. However, a further receptor would be required to allow neuronal infection. By screening a phage antibody library for antibodies which block MV infection of human neuronal cells we have identified a putative receptor (molecule X) in neuronal cells.When non-permissive Vero cells were transfected to express this molecule they became susceptible to WT MV. We have also carried out immunostaining in MV infected and non-infected regions of human brain tissue. Molecule X is highly expressed in the human CNS. We are currently investigating if this molecule can also be used as a receptor by CDV. If both PVRL4 and receptor X are used by CDV for CNS infection this will raise major concerns for possible human infection with this highly neurovirulent veterinary virus

Kavita kakkar

Sanjay Gandhi Post Graduate Institute of Medical Sciences, India

Title: Impact of A3B deletion polymorphism on HIV-1 susceptibility

Time : 14:45-15:05

Biography:

Kavita Kakkar hails from the sacred city of Allahabad and is a PhD scholar. She completed her B.Sc (Hons) from Allahabad Agricultural Institute Deemed University and M.Sc from Vellore Institute of Technology. Her keen and enthusiastic approach towards the field of Microbiology has motivated her to attend multiple academic programs, conferences like ASICON etc. and fetched her an award during BIOXPLORE’07. Her five year stint with Sanjay Gandhi Post Graduate institute of Medical Sciences, Lucknow where she currently working as SRF on an ICMR project on HIV, has further sharpened her aptitude for research and thirst for knowledge. She is currently pursuing Ph.D on aspect of HIV pathogenesis, and have 2 publications and 6 nucleotide submissions. She strives to maintain a good learning curve throughout her career.

Abstract:

The human APOBEC3 (A3) family proteins of cytidine deaminases potently restricts HIV-1 replication (A3B), a gene involved in innate immunity exhibiting insertion–deletion polymorphism across world population and provides intrinsic immunity to retroviral infection. In humans, a high-frequency distribution of 29.5kb deletion occurs between exon 5 and exon 8, and A3B deletion genotype is observed. This deletion results in complete loss of A3B coding region. The present study investigated the effect of insertion(I)/deletion(D) polymorphism frequencies of A3B gene on susceptibility to HIV infection among 84 HIV seronegative (HSN) and 26 HIV seropositive (HSP) individuals in North Indian population, which was assessed based on frequencies of three genotypes: deletion-homozygous (D/D), hemizygous (D/I), and no deletion (I/I) genotypes between infected and uninfected cohorts. The genotypic analysis showed no significant difference in the ratios of A3B genotype between HIV infected (I/I 57.7%, I/D 30.8% and D/D 11.5%) and HIV uninfected ( I/I 42.9%, I/D 30.9% and D/D 26.2%) individuals and also no association of deletion allele was seen on HIV disease susceptibility among HSN (I vs D odds ratio = 0.327, P value 0.093, 95% CI= 0.085 and I vs I/D odds ratio = 0.738, P value = 0.550, 95% CI = 0.273) when compared with HSP. These results suggest no significant effect of A3B gene polymorphism on the HIV-1 susceptibility and also no association was found with hemizygote genotype. Hence, preliminary analysis of the data has shown that there is no significant impact of A3B deletion on HIV-1 susceptibility.

Biography:

Education: • Enrolled in PhD, Biochemistry from Ziauddin University Karachi • M.Phil Biochemistry from Ziauddin Medical University Karachi-Pakistan in collaboration with HEJ, International Research Institute for Chemical Sciences, University of Karachi in May, 2006 • M.B.B.S from King Edward Medical College Lahore in the year 1996 • B.Sc from Punjab University in the year 1992 • F.Sc from Multan in the year 1989 • S.Sc from Multan in the year 1987 Research Experience : Working on PCR, Electrophoresis, Sequencing , Bioinformatics and Phylogenetics from April 2012 to date • Five years and six months Research experience in Department of Biochemistry at Ziauddin Medical University and HEJ Research Institute, International Centre of Chemical Sciences in collaboration as research student from 1st March 2001 – 31st May, 2006.

Abstract:

Background The human papillomavirus (HPV) has been evolved as a new culprit of malignant and pre malignant oral lesions of tobacco chewers in several studies . Taking into account this role of HPV in oral mucosa, the objective of this study was to detect the presence of HPV and its genotypes 16, 18 in different lesions of oral cavity of tobacco chewers. Methods A 15-30 ml of buccal wash sample was collected from 522 subjects (440 males and 82 females) with leukoplakia, erythroplakia, submucous fibrosis and oral squamous cell carcinoma after an informed consent. Gentle brushings from the lesions were taken from subjects with the help of a brush at the other end of dental floss and the buccal wash was stored at 4°C until DNA extraction. DNA was extracted and PCR was performed using HPV consensus primers Gp5+/Gp6+. The genotyping for HPV 16, 18 was performed using HPV 16, 18 specific primers. Results The 84% (440/522) of males were affected by oral premalignant lesions whereas, submucous fibrosis was found to be the most frequent pre malignant oral lesion (192) out of total 522 cases. Out of 192 SMF cases, 82 were HPV positive with 91% (75) males and 9% (7) females. In SMF, 74% (61) have genotypes other than HPV 16 and 18. HPV 16 and 18 were frequent in 20 patients with OSCC. HPV 18 was frequent genotype in patients with erythroplakia in this study. Conclusion The patients with SMF are at greater risk of having HPV. HPV 16 and 18 were more frequent malignant genotypes but the possibility of other HPV genotypes causing pre malignant and malignant lesions requires further investigation.

  • Track 4: HIV and Other Retroviral Diseases Affecting Afro-Asian Continents
Location: San Antonio, USA

Session Introduction

Arun Rajasekaran

Kasturba Medical College, India

Title: APOL1 risk variants predict histopathology and progression to ESRD in HIV-related kidney disease

Time : 13:05-13:25

Biography:

Arun Rajasekaran is working as a faculty member in Kasturba Medical College, India.

Abstract:

With earlier institution of antiretroviral therapy, kidney diseases other than HIV-associated nephropathy (HIVAN) predominate in HIV-infected persons. Outcomes for these diseases are typically worse among those infected with HIV, but the reasons for this are not clear. Here, we examined the role of APOL1 risk variants in predicting renal histopathology and progression to ESRD in 98 HIV-infected African Americans with non-HIVAN kidney disease on biopsy. We used survival analysis to determine time to ESRD associated with APOL1 genotype. Among the 29 patients with two APOL1 risk alleles, the majority (76%) had FSGS and 10% had hypertensive nephrosclerosis. In contrast, among the 54 patients with one APOL1 risk allele, 47%had immune-complexGNas the predominant lesion and only 23%had FSGS. Among the 25 patients with no APOL1 risk allele, 40%had immune-complex GN and 12%had FSGS. In 310 person-years of observation, 29 patients progressed to ESRD. In adjusted analyses, individuals with two APOL1 risk alleles had a nearly three-fold higher risk for ESRD compared with those with one or zero risk alleles (P=0.03). In summary, these data demonstrate an association between APOL1 variants and renal outcomes in non-HIVAN kidney disease, suggesting a possible use for APOL1 genotyping to help guide the care of HIV-infected patients.

Biography:

Edith Adanna Onwuliri holds a B. Pharm Degree in Pharmacy and an M.Sc Degree in Applied Microbiology from the University of Jos, Plateau State Nigeria. She just concluded a Ph.D programme at the University of Nigeria, Nsukka in the Department of Pharmaceutics. She is a lecturer in the Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, University of Jos, Nigeria. She is interested in therapeutic investigations of local plants for their antimicrobial effects and their formulation into suitable dosage forms. She has 11 publications in various journals. Dr. Mrs is married to Professor F.C. Onwuliri and their marriage is blessed with six children.

Abstract:

The study evaluated the CD4+ T- Cells of HIV/AIDS patients who enrolled for Highly active Antiretroviral Therapy (HAART) between the periods January to August, 2009 at Federal medical centreKeffi-Nasarawa state. Five hundred confirmed HIV patients enrolled for the programme while twenty (20) patients who were not yet enrolled for the programme were monitored as control group. Out of the 500 patients enrolled for the programme sixty – five (65) patients were lost to follow – up. The remaining four hundred and thirty five (435) Patients blood samples were collected and analysed using Becton – Dickinsoncyflowcytometer both at baseline and at sixth month of HAART. The number of female patients enrolled for the programme were 303(70%) while male patients were 132(30%). The result showed that three hundred and sixty-seven (367) (84%) patients, had increase CD4+ T cell count while sixty – eight, 68 (16%) patients failed to respond to treatment. The mean CD4+ cell count of patients at baseline for various age groups were, 393 cells/μl for age 20 – 30 years; 191 cells/μl for 31 – 40 years, 195 cells/μl for 41 – 50 years and 297 cells/μl for 51 years and above. At sixth month of HAART, patients mean CD4+ T. cell count increased as follows; age 20 – 30 years 623 cells/μl, 31 – 40 years 343 cells/μl, 41 – 50 years 338 cells/μl and 51 years above 381 cells/μl. The age group 20 – 30 years showed the highest number of patients with CD4 + cell increase by 260(60%), 31 – 40 years 126(29%), 41 – 50 years 38(8%), 51 years and above 11(3%.) From the result, it implies that when patients are placed on good Antiretroviral Therapy, there is significant improvement in their immune system which however, decreases with age. Further studies may be required in HIV patients with baseline CD4+ cell count >400 cells/μl before commencement of therapy as there was no significant improvement in their CD4+ cell count after six months of HAART.

Biography:

Herve Perron has completed his PhD in 1991 (Virology) and HDR (Professor Thesis; Biology) in 2000 from the Faculty of Medicine, Joseph Fourier University, Grenoble-France. He isolated and characterized a novel retroviral element from Multiple Sclerosis (MSRV), itself defining a novel family of human endogenous elements (HERV-W). He reviewed about a hundred manuscripts for more than 40 scientific and medical journals and is author in about 70 peer-reviewed publications. He is presently Chief Scientific Officer of Geneuro SA, Geneva-Switzerland. Geneuro develops innovative humanized antibody treatments for Multiple Sclerosis and for other diseases involving HERV-W as psychoses associating neuroinflammation.

Abstract:

Human endogenous retroviruses (HERV) are complex and heterogeneous multicopy families of genetic elements which represent about 8 % of the human genome. HERV copies can retain transcriptional activity but complete and functional provirus have not been identified so far. Retrovirus-like particles with reverse transcriptase (RT) activity were shown in macrophage cultures from patients with multiple sclerosis (MS). PCR extension to all retroviral genes in purified particles with specific buoyant density and reverse transcriptase activity was achieved and unravelled the previously unknown HERV-W family. As many unfixed HERVs (Marchi et al. J.Virol. 2014), the corresponding provirus has not already been isolated. Nonetheless, HERV-W env, gag and pol-encoded proteins are detected in all active MS brain lesions analysed to date, whereas HERV-W envelope displays potent neuroinflammatory pathogenicity. We have now identified cultured cells expressing all HERV-W gag-pol-env encoded proteins detected with specific monoclonal and polyclonal antibodies by WB analysis, along with RT activity in supernatants. Polyproteins, cleaved capsid and RT bands are seen for gag and pol, as full-length glycosylated monomer and multimers for env, indicating that HERV-W copies with complete orfs for gag, pol and/or env exist in human cells but are not described in the existing databases. We should therefore seek for still undescribed and unfixed HERV-W copies in DNA. Analysing their distribution in the human populations as in patients with diseases for which HERV-W proteins and RNA levels are associated with etiopathogeny or lesions, could clarify the involvement of HERV-W elements in several chronic inflammatory diseases.

Speaker
Biography:

Dr. Brown completed her Ph.D from the Albert Einstein College of Medicine of Yeshiva, University and postdoctoral studies at the Aaron Diamond AIDS Research Center. She is the Director of the Johns Hopkins Internship in Brain Science Program, Co-Director of the Developmental Core for The NIMH Center for Novel Therapeutics for HIV-Associated Cognitive Disorders, Co-Director of Translational Research in NeuroAIDS and Mental Health and a Fellow of Keystone Symposia on Molecular and Cellular Biology 2014. Dr. Brown has published 17 papers in reputed journals and serves as Managing Editor for the Frontiers in Bioscience Special Issue Series.

Abstract:

More than 30 years into the HIV epidemic, significant worldwide morbidity and mortality due to infection with this virus continues despite the increased availability of combination anti-HIV therapies, which can effectively suppress viral replication. HIV enters the central nervous system early after penetrating mucosal portals and disseminating where it infects and replicates robustly in resident microglia and macrophages. Neurons do not serve as hosts for HIV and the injury and dysfunction seen in individuals with HIV-associated cognitive impairment occurs through a network of indirect toxic mechanisms, many of which remain to be defined. Cellular activation of multiple cell types in the brain, as a result of HIV infection, leads to levels of inflammation that are in excess of that seen under normal homeostatic conditions. We have been investigating the role of the proinflammatory cytokine osteopontin in the pathogenesis of HIV-associated neurocognitive disorders (HAND). Osteopontin is an early immune activation marker implicated in the regulation of T-lymphocyte function and balance between cell- and humoral immunity. In macrophages, OPN can regulate cell migration and survival and is elevated in individuals with HAND. We have used a combination of molecular and cellular approaches including next generation high-throughput sequencing (RNA-Seq), surrogate culture models to define the domains of OPN required for its ability to enhance HIV replication, and ex vivo studies using biological samples from HIV-infected individuals with or without cognitive impairment and controls to delineate its role in HIV-related neuropathogenesis. By understanding how HIV commanders macrophage signaling networks, we may be able to develop novel therapeutic strategies to counteract the pathogenic effects of the virus.

Speaker
Biography:

Neal S. Rote completed his Ph.D. at Temple University School of Medicine and postdoctoral studies at Heidelberg University and UCLA School of Medicine. He is the William Weir, M.D. Professor of Reproductive Biology and Professor of Pathology at Case Western Reserve University School of Medicine and Academic Vice Chair and Director of Research in the Department of Obstetrics and Gynecology, University Hospitals Case Medical Center, Cleveland, OH. He has published more than 110 papers in reproductive biology and 75 chapters and books, been NIH-funded for 32 years, and served on many NIH review committees.

Abstract:

Normal human placentation requires differentiation of specialized cells (villous cytotrophoblast into syncytiotrophoblast); principally characterized by intercellular fusion and production of the hormone chorionic gonadotropin (hCG). Concurrently trophoblasts express cellular genes of apparent retroviral origin (endogenous retroviruses; ERV), including endogenous retroviral element ERV3 env. We reported that, unlike other retroviral env regions that encode fusion proteins, ERV3 env regulated induction of the β subunit of hCG (β-hCG). The biological relevance of ERV3 env was questioned in a report of adults with homozygous stop mutations leading to a “natural knockout” of ERV3 env although a truncated (p25) SU protein was produced that lacked the biologically active regions typically attributed to exogenous or endogenous retroviral Env proteins. The p25 region was never tested for capacity to induce β-hCG. ERV3 env contains two proposed translational start sites at nt 595 and nt 715. We cloned and inserted the entire ERV3 env open reading frame (ERV3), the SU region, and the p25 region, beginning at both proposed start sites, in stable expression vectors into BeWo cells (a model for villous cytotrophoblast differentiation when treated with forskolin), quantified levels of intracellular β-hCG and actin by western blot analysis, and data expressed as means (SD) of the ratio of β-hCG to actin of three independent experiments. β-hCG was not detectable in untreated BeWo or those transfected with vector alone (negative controls) and maximally expressed in forskolin-treated cells (positive control; 1.83 + 0.83). Transfection with ERV3, ERV3 SU, and p25 induced significantly (P<0.01) greater levels of β-hCG expression. BeWo cells were also stably transfected with vectors expressing siRNAs targeted to regions near the start sites, and the cell lines treated with forskolin or vehicle alone for 24, 48, and 72 hr. Transfection with si670, targeted to nt 670-688, between the proposed start sites, completely prevented induction of β-hCG by forskolin at all time points. Thus, ERV3 env is an atypical retroviral element with a unique trophoblast hormone regulatory site in the SU region and in the p25 truncated protein expressed in individuals with described homozygous stop mutations.

Speaker
Biography:

Dr. Sholukh, a native of Belarus, completed his PhD in 2002. His PhD work was devoted to study the regulation of intracellular signaling and transduction of light signals in the retinal photoreceptor. Since 2004 Dr. Sholukh’s research interests have broadened to immunology and antibody engineering. His work also included the discovery of an important soluble CXC chemokine receptor 2. In 2009, Dr. Sholukh joined the laboratory of Dr. Ruprecht at the Dana-Faber Cancer Institute (Harvard Medical School) and in 2013 he moved to Texas Biomedical Research Institute. His current interests are focused on dissecting the humoral immune response to HIV and HIV vaccine candidates as well as on the vaccine design.

Abstract:

An ultimate goal for HIV vaccine design is the induction of cross-reactive neutralizing antibodies (nAbs). The exceptional diversity of HIV makes it impossible for any AIDS vaccine recipient to be exposed to virus strains exactly matching the immunogen given. For that reason, heterologous SHIV challenges are essential for realistic vaccine efficacy testing in primates. We sought to test whether passive immunization with SHIVIG, defined as polyclonal IgG raised in rhesus monkeys (RMs) with chronic clade C SHIV infection, could protect against multiple low-dose intrarectal challenges with a tier 2 SHIV-2873Nip carrying an HIV-C envelope that was heterologous to any viruses or envelopes against which the IgG responses had been elicited. SHIVIG demonstrated binding to HIV Gag, Tat and Env of different clades and competed with the broadly neutralizing antibodies b12, VRC01, 4E10, and 17b. SHIVIG neutralized tier 1 and tier 2 viruses, including SHIV-2873Nip the heterologous challenge virus. NK-cell depletion decreased the neutralizing activity of SHIVIG 20-fold in PBMC assays. Although SHIVIG neutralized SHIV-2873Nip in vitro, this polyclonal IgG preparation failed to prevent acquisition after repeated intrarectal low-dose virus challenges, but at a dose of 400 mg/kg, it significantly lowered peak viremia (P=0.001). Surprisingly, single-genome analysis revealed a higher number of transmitted variants at the low dose of 25 mg/kg, implying increased acquisition at low SHIVIG levels. In vitro, SHIVIG demonstrated complement-mediated Ab-dependent enhancement of infection (C’-ADE) at concentrations similar to those observed in plasmas of RMs treated with 25 mg/kg of SHIVIG at the time of virus exposure. These primate model data suggest a dual role for polyclonal anti-HIV-1 Abs depending on plasma levels upon virus encounter.

Eduard Karamov

Ivanovsky Institute of Virology
Russia

Title: Molecular Genetics of HIV-1 Strains Spreading in Russia

Time : 14:25-14:45

Biography:

Eduard Karamov is working as Head of Laboratory of Immunochemistry in Ivanovsky Institute of Virology.Professor Eduard Karamov is in Retrovirology research since 1985. The first Russian HIV strain was isolated in his laboratory. He supervised the development of the first Soviet diagnostic tests for HIV and antivirals for AIDS treatment. He published over 100 articles and made over 100 presentations at international meetings. He supervised 15 Ph.D. and Dr.Sci. Fellowships. He is a member of Editorial Board of international and Russian scientific journals

Abstract:

Introduction HIV-1 epidemic in Russia is dominated by a monophyletic subtype A1 variant, derived from a strain originating in Central Africa, which began to spread among IDUs in 1995. The generalized epidemic observed in Russia affects populations at risk (IDUs, CSWs, and MSM), with rates of HIV-1 incidence being among the highest in the world. The number of HIV-1 cases recorded in Russia has exceeded 800 000, but experts estimate that the total amount of the infected individuals is about 2 000 000 (> 1% of the population of Russia). Objectives To analyze phylogenetic clustering and predicted coreceptor usage in Russian HIV-1 sequences. Methods Plasma samples were collected in the Moscow area (13 samples, 2010) and the Perm area (10 samples, 2011). RNA was extracted from paper spots and directly amplified by RT-PCR. Sequences of PR-RT and V3 were obtained for 14 and 17 samples, respectively; full-length genome sequencing was performed for two samples. Phylogenetic sequence analysis was done via maximum likelihood with RAxML. Genotypic prediction of coreceptor usage was done by V3 net charge and the online programs Geno2Pheno and PSSM. Results Five samples from the Perm area, sequenced in PR-RT, were A1. Among the nine samples from the Moscow area, sequenced in this segment, six were A1, two – CRF02_AG, and one belonged to subtype B. Full-length genome sequencing of the two samples from the Moscow area demonstrated that one of them was A1 and the other – CRF02_AG. CCR5 coreceptor usage was predicted by all methods in all the 17 V3 sequences (nine samples were isolated in the Perm area and eight – in the Moscow area). Conclusions Molecular-epidemiological studies demonstrated that the nascent HIV-1 epidemic was associated with the introduction of diverse viruses in distinct risk groups. The rapid spread of the homogenous IDU-A strain resulted in this virus’ becoming predominant across Russia and most Eastern European and Central Asian countries of the former Soviet Union. It is obvious that the social and economic deterioration and the massive rise in drug abuse did play a role in this explosive epidemic. The new stage (generalized epidemic) is characterized by active propagation of HIV-1 subtype A1 and multiple outbreaks of CRF02_AG infection. The latter strain is similar in sequence to isolates from Uzbekistan and Kyrgyzstan, rather than African or Western European, which is likely related to labor migration. This is the first report of the circulation of CRF02_AG in the Moscow area. Of interest, double recombinants A/AG have already been detected in Russia and Kyrgyzstan. It is also intriguing that none of the sequences of this study contained major drug resistance mutations.

Speaker
Biography:

Dr Gokul C Das is currently a member of the faculty in the Department of Medicine, of the Center for AIDS research, Center for Drug Discovery and of the Dan L Duncan Cancer Center at the Baylor College of Medicine (BCM), Houston. Previously,he was a professor of molecular biology at the University of Texas Health Science Center at Tyler (UTHSCT). Dr Das had his Ph.D. degree in Biophysics from the University of Kolkata, India. After his postdoctoral work at the Institute de Biologie Moleculaire et Cellulaire du CNRS, Strasbourg, France and at the Oak Ridge National Laboratory, Oak Ridge, TN, he was a visiting scientist at NIH working on DNA tumor viruses. He continued his interest as a Professor of Molecular Biology at UTHCT. His Current interest is to understand molecular pathways in pathogenesis induced by Hepatitis C Virus (HCV) and HIV, or in HCV-HIV co-infection, and to use both a cellular and humanized animal model to develop antiviral strategies. Dr Das is currently on the editorial board of a number of international journals. He was the receipient of international guest scientist awards (1997,2000) from the Ministry of Science and Technology, Japan and was a member of the biotechnology delegation to China. .

Abstract:

Hepatitis C virus (HCV) infection is a global health care problem affecting over 200 million individuals without any effective therapy and vaccine. In addition to a spectrum of liver diseases, chronic infection induces insulin resistance (IR) leading to metabolic syndrome and type 2 diabetes (T2DM) by dysregulating insulin signaling pathway(s). Development of IR not only accelerates the progression of liver disease, but also makes IFN-based therapy less-responsive. Thus understanding of the molecular basis of IR and non- response to therapy is pivotal for novel therapeutic development. Our central hypothesis is that both insulin receptor substrates (IRS-1 and IRS-2) are involved in dysregulation of insulin signaling by two entirely different mechanisms, the former through Ser 312 phosphorylation and the latter through down regulation by mi RNAs. Our major findings are: a) metabolic pathways for glucose homeostasis, insulin signaling and autophagy are dysregulated and they interact to contribute to IR, b) autophagy inhibitor Beclin-1 and energy sensors AMPK and mTOR are hyper- phosphorylated which is inhibited by IFNα, c) pathways leading to IR is activated within two weeks after infection, d) has-let 7 family of miRNAs are upregulated and target IRS-2, but these are down regulated in IFN treated cells. Our results suggest that strategies aimed at inactivating multiple pathways combinatorial effect of down regulating IRS-1 pathways and miRNA regulation of IRS-2 may lead to a better therapeutic outcome

Biography:

Yuetsu Tanaka is working as faculty in University of the Ryukyus, Japan.His field of research is virology and immunology.

Abstract:

The number of human T cell leukemia virus type-1 (HTLV-1)-infected individuals in the worldhas been estimated to be ~20 million. However, no prophylaxis vaccines or drugs against HTLV-1 infection have been developed. Thus, for establishing the basis of protective vaccines against HTLV-1,we have humanized a rat monoclonal anti-HTLV-1-neutralizing antibody, LAT-27, that recognizes the HTLV-1 gp46 amino acids 191-196. The humanized LAT-27 (hu-LAT-27) completely blockedboth HTLV-1-infection in vitro at a concentration of 5 microgram/ml as determined by syncytium-formation and transformation inhibition assays. A51Cr-releace assay showed that hu-LAT-27 efficiently lysed HTLV-1-infected cells by ADCC in the presence ofautologous or allogeneic fresh PBMCs. Magnetic cell depletion assays showed that the main effector cells involved in the ADCC were NK cells. The ADCC activity of hu-LAT-27 (human IgG1) was superior to that of original LAT-27 (rat IgG2b). When hu-PBL-SCID mice were pre-infused i.v.with hu-LAT-27, but not a control chimeric human antibody, all the mice were completely protected against HTLV-1 infection. These results indicate that this new humanized anti-HTLV-1 antibody is potent not only in blocking of new infection of human T cells with HTLV-1, but also in surveillance of HTLV-1-infected cells, indicating a possible potential of hu-LAT-27 for passive immunization against HTLV-1 infection.

Gulnara A. Yuldasheva

Scientific Centre for Anti-Infectious Drugs
Kazakhstan.

Title: A Quantum-Chemical Model of the Inhibition of HIV-1 Integrase Action by Molecular Iodine.

Time : 14:45-15:05

Biography:

Gulnara A. Yuldasheva is working in Scientific Centre for Anti-Infectious Drugs Kazakhstan.

Abstract:

A distinctive feature of the drugs having anti-HIV and anti-viral action (AHD) [1-2] is that they contain not only an iodine-polymer complex, but also lithium and potassium halogenides. Using X-ray data for iodine--dextrin complexes and the results of quantum-chemical ab initio RHF/3-21G** level calculations a model of the active complex (AC) of AHD was proposed. It is suggested that the drug active complex contains molecular iodine located inside the -dextrin helix and coordinated by lithium halogenides and polypeptides. Electronic structure of I2 in this complex differs from its characteristics in complexes with organic ligands or the free I2. In the AC under study the molecular iodine displays the acceptor (donor) properties towards polypeptides (lithium halogenides) [1]. In paper [3] UV- and IR-spectroscopy has been used to study the water-glycine - KI3 - LiCl –ethanol system that forms AC of AHD. It has been shown that in this system conditions are created for the formation of an iodine complex compound, in which the molecular iodine reveals the acceptor properties towards glycine, and the donor properties towards the LiCl-ethanol complex. A mechanism of AHD anti-HIV action has been proposed. Under the influence of molecular iodine contained in the AC of AHD the structure of HIV DNA is modified: the nucleotides of the viral DNA that are more π-donor-active than peptides form a stable complex with molecular iodine and lithium halogenides [1]. Using UV- and IR-spectroscopy and quantum-chemical DFT/B3PW9 we have confirmed the existence of the molecular iodine complex coordinated by lithium halogenides with nucleotide (nucleotideI2 LiCl) in the system containing the AGA nucleotides triplet and the AC complex of AHD. The interaction of molecular iodine coordinated by lithium halogenides with the viral DNA and the HIV-1 integrase co-factor has been studied by DFT/B3PW9 method. Calculations have shown that complex nucleotideI2 LiCl may prevent the active catalytic fragment of HIV-1 integrase from interacting with the virus DNA. Complex nucleotideI2 LiCl may become the center of another nucleoprotein complex in which molecular iodine interacts both with the virus DNS and the active catalytic domain of HIV-1 integrase Experimental data on the anti-HIV effect of AHD [2] and the results of calculations suggest that the molecular iodine coordinated by lithium halogenides can be regarded as a compound inhibiting the catalytic center of integrase. Reference. 1. G.A Yuldasheva, G.M. Zhidomirov, A.I. Ilin (2012) Biotechnology and Appled Biochemistry 59(1):29-34. 2. International Application (2000) Ilyin Alexandr, Gab-rielyan Emil, Mkhitaryan, Levon Antiviral and antibacte-rial pharmaceutical preparation “Armenicum” And its use for treatment of infectious diseases. Patent No.: PCT/AM 2000/000002. 3. G.A. Yuldasheva, G.M. Zhidomirov, J. Leszczynski, A.I. Ilin (2013) J. Mol. Struct. 1033: 321–330.

Vladimir Zajac

Slovak Academy of Sciences, Slovakia.

Title: Intestinal and Respiratory Tract Bacteria and Yeasts in pathogenesis of AIDS

Time : 11:45-12:05

Speaker
Biography:

Vladimir Zajac has completed his PhD. in 1982 at the Cancer Research Institute of Slovak Academy of Sciences in Bratislava (Slovakia), where he was from 1996 to 2010 the head of Department of Cancer Genetics. He joined the Medical Faculty of the Comenius University as Associate Professor of Genetics in 2008. He has published 59 papers mostly in reputed journals and he was editor of the book „Bacteria, viruses and parasites in AIDS process“ (InTech, 2011).

Abstract:

Recently accumulate the records, which show that the main site of HIV infection and CD4+ T cell loss is in the GIT and the other mucosal tissue rather than in blood. The HIV-1 was also detected in bowel crypt cells and the lamina propria. It has also been proven that various forms of HIV reservoirs persist in practically all patients receiving HAART. Persistence of HIV was also detected in GALT despite HAART. These facts indicate that in the pathogenesis of AIDS may come into play additional factors. Due to the fact that on mucosa of the colon is a large variety of bacteria and yeast, it is necessary to follow their role in process of pathogenesis. In accordance to this idea, we tested the bacteria and yeast in Slovak and American AIDS patients and HIV-positive children in Kenya and Cambodia for the presence of HIV-like sequences. Using specific primers have been in DNA of these bacteria and yeast identified sequences with high homology to HIV-1. Expression of these sequences has been demonstrated with monoclonal antibodies against p17, p24, p55, gp41 and gp120. Based on these results the presence of HIV-like sequences in bacteria of the patients may be hypothetically explained as follows: 1) bacteria and yeasts are a natural host of HIV sequences; 2) these sequences were transferred into intestinal bacteria from degraded human cells. Hypothetically HIV sequences can be in bacteria and yeast intestinal and respiratory tract present since the beginning of mankind. Thanks to countless epidemics that accompany humanity since its inception, microbes carry these sequences have been largely removed. Administration of antibiotics, drugs and homo and anal sex has recently propagated again. In this way, pathogenic microbes, especially drug resistant, which used to be in the minority moved into the majority. Microbes bearing HIV-like genetic information penetrate from the intestinal tract into the blood. Because of their affinity to lymphocytes, infected or lysed them and a process of immunodeficiency started

  • Track5: Organ Specific Cancers and Tumour Virology
Location: San Antonio, USA

Session Introduction

Felipe Samaniego

University of Texas MD Anderson Cancer Center, USA

Title: Human herpesvirus 8 K1-mediated inhibition of fas-mediated apoptosis provides a new insight into regulation of fas signaling

Time : 16:40:17:00

Biography:

He is working as Associate Professor, Department of Lymphoma/Myeloma, Division of Cancer Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX Academic Appointments Assistant Professor, Department of Immunology, University of Texas M.D. Anderson Cancer Center, Houston, TX, 2003-2006 Assistant Professor, Human Virology, Institute of Human Virology, University of Maryland Medical Center, Maryland, MD, 1996-1999 Honors and Awards 2011 US News and World Report Top Docs 2002 AACR-MICR Council Member, AACR-MICR 1989 Daland Fellowship, Alternate Fellow 1989 Robert Wood Johnson Foundation Fellow

Abstract:

Kaposi’s sarcoma and primary effusion lymphomahave been linked to human herpesvirus 8 infection. Key to the pathogenesis of these cancers and the persistence of the virus is the transmembrane proteinK1, which induces lymphoproliferation and immortalization of lymphocytes throughunknown mechanisms. K1 transgenic mice showed expansion of the lymphoid system; enlarged spleens, lymph nodesnodes, and development of lymphoma and sarcoma tumors. The K1 splenocytes were resistant to apoptosis induced by agonistic anti-Fas antibody. We demonstrated that K1-mediatedinhibition of Fas agonistinducedcaspase8- dependent apoptosis in transgenic mice did not require an immunoreceptor tyrosine-based activation motif (ITAM) of K1. K1 bound Fas through the immunoglobulin-like extracellular domain, and blocked apoptotic signaling via interference with the binding of FasLand subsequently suppressed formation of the death-inducing signaling complex. We anticipated that the K1 mimics the function of endogenous proteins participating in regulation of Fas receptor signaling. To identify these proteins, we purified activation-resistant Fas protein complexes and identified nucleolin as protein associated with activation-resistant Fas. To confirm Fas-regulatory function of nucleolin, we over-expressed nucleolin in mouse liver where it blocked Fas-mediated apoptosis. On the other hand, nucleolin knockdown in cells enhanced the levels of Fas-mediated apoptosis by enhancing the binding of Fas ligand. These results are consistent with the role of other Fas receptorbinding proteins such as hepatocyte growth factor receptor and CD44that bind Fas and interfere with the initiation of Fasmediated apoptosis. These data reveal a novel mechanism of regulation of Fas-mediated apoptosis by a viral receptor-like protein and shed light on the other similar regulators of Fas apoptotic signaling.

Biography:

A. Gritzapis was granted a PhD at the age of 27 from the Department of Biological Chemistry in the Medical School of the University of Ioannina, Greece and was employed until 2012 as principal investigator at the Cancer Immunology Immunotherapy Center of the Department of Immunology at Saint Savas Cancer Hospital, Athens Greece. He is currently employed as principal investigator in the department of Immunology and Cellular Biology at Locus Medicus S.A., Athens, Greece. He has published more than 50 papers in reputed journals.

Abstract:

Viral presence in sperm is a cause of infertility. Viral factors as a cause of male subfertility have not greatly concerned the medical community up until now. On the contrary, a correlation between high numbers of Natural Killer lymphocytes (ΝΚ) in the blood of women with a history of subfertility and/or miscarriages, and the presence of subclinical herpes viremia (HSV1-2, EBV, CMV, HHV6 and HHV7) has been described. Observation of miscarriage material revealed that NK mostly infiltrated at the implantation site, while the blood NK levels in a portion of these women were normal. This can be explained if the embryos in these cases were by themselves antigenic due to the presence of viral (at least herpes-viral) antigens, originating from the male through the sperm cells, including spermatozoa. These antigens would be expressed and presented to the woman's immune system by fetal cells causing the NK response. A method of intracellular detection of pathogens (viruses and Chlamydia trachomatis) has been developed. The sperm cells are fixed, permeabilized and DNA digestion is accomplished with DNase I. Incubation with antibody against each pathogen is followed by incubation with a fluorescent conjugated secondary antibody. The samples are acquired in a flow cytometry apparatus and analyzed with suitable software. Results so far, show that a significant percentage of samples taken from infertile men were found to be infected by intracellular chlamydia and/ or viruses. Especially in men infected with Chlamydia trachomatis, microbial load fell or the infection disappeared following antibiotic treatment. Furthermore, they also improve their Teratozoospermia Index (TZI) and the mean value of abnormalities per spermatozoon.

A D Gritzapis

LOCUS MEDICUS S.A, Greece

Title: Intracellular Detection of Infectious Pathogens in Sperm Cells

Time : 17:40-18:00

Biography:

A. Gritzapis was granted a PhD at the age of 27 from the Department of Biological Chemistry in the Medical School of the University of Ioannina, Greece and was employed until 2012 as principal investigator at the Cancer Immunology Immunotherapy Center of the Department of Immunology at Saint Savas Cancer Hospital, Athens Greece. He is currently employed as principal investigator in the department of Immunology and Cellular Biology at Locus Medicus S.A., Athens, Greece. He has published more than 50 papers in reputed journals.

Abstract:

Viral presence in sperm is a cause of infertility. Viral factors as a cause of male subfertility have not greatly concerned the medical community up until now. On the contrary, a correlation between high numbers of Natural Killer lymphocytes (ΝΚ) in the blood of women with a history of subfertility and/or miscarriages, and the presence of subclinical herpes viremia (HSV1-2, EBV, CMV, HHV6 and HHV7) has been described. Observation of miscarriage material revealed that NK mostly infiltrated at the implantation site, while the blood NK levels in a portion of these women were normal. This can be explained if the embryos in these cases were by themselves antigenic due to the presence of viral (at least herpes-viral) antigens, originating from the male through the sperm cells, including spermatozoa. These antigens would be expressed and presented to the woman's immune system by fetal cells causing the NK response. A method of intracellular detection of pathogens (viruses and Chlamydia trachomatis) has been developed. The sperm cells are fixed, permeabilized and DNA digestion is accomplished with DNase I. Incubation with antibody against each pathogen is followed by incubation with a fluorescent conjugated secondary antibody. The samples are acquired in a flow cytometry apparatus and analyzed with suitable software. Results so far, show that a significant percentage of samples taken from infertile men were found to be infected by intracellular chlamydia and/or viruses. Especially in men infected with Chlamydia trachomatis, microbial load fell or the infection disappeared following antibiotic treatment. Furthermore, they also improve their Teratozoospermia Index (TZI) and the mean value of abnormalities per spermatozoon.

Speaker
Biography:

High-risk human papillomaviruses (most frequently HPV-16) are sexually transmitted infections responsible for pre- and cancerous lesions of cervix as well as a high percentage of anal, vaginal, vulvar, penile and oropharyngeal cancers. Effective vaccines based on the HPV core protein L1 currently approved for prevention of new HPV infections in young adults are ineffective to treat existing infections/lesions which predominantly express the viral E6 and E7 oncogenes but not L1. Therapeutic vaccines based on the E6 and E7 tumor antigens are urgently needed, especially for people in the developing world to combat HPV diseases. We discovered specific HPV-16 E6 and E7 peptides for which memory T cell immunity is associated with recurrence-free survival in women treated for HPV pre-cancerous cervical lesions. In mouse model studies, mucosal intranasal immunization employing these E6 and E7 peptides admixed with novel adjuvants induced HPV-specific systemic and mucosal effector CD4/CD8 T cell responses that prevented HPV+ tumor formation. Therapeutic mucosal intranasal vaccination with these peptides in combination with adjuvants significantly reduced HPV tumor growth in mice affording long term survival advantage. Furthermore, combination of intranasal vaccination with the E6 and E7 peptides with immune check point blockade antibody treatmentresulted in significant tumor regression leading to eradication. These data support future clinicaltesting of combination vaccine immunotherapy strategies for effective treatment and possibly eradication of pre- and cancerous HPV lesions.

Abstract:

High-risk human papillomaviruses (most frequently HPV-16) are sexually transmitted infections responsible for pre- and cancerous lesions of cervix as well as a high percentage of anal, vaginal, vulvar, penile and oropharyngeal cancers. Effective vaccines based on the HPV core protein L1 currently approved for prevention of new HPV infections in young adults are ineffective to treat existing infections/lesions which predominantly express the viral E6 and E7 oncogenes but not L1. Therapeutic vaccines based on the E6 and E7 tumor antigens are urgently needed, especially for people in the developing world to combat HPV diseases. We discovered specific HPV-16 E6 and E7 peptides for which memory T cell immunity is associated with recurrence-free survival in women treated for HPV pre-cancerous cervical lesions. In mouse model studies, mucosal intranasal immunization employing these E6 and E7 peptides admixed with novel adjuvants induced HPV-specific systemic and mucosal effector CD4/CD8 T cell responses that prevented HPV+ tumor formation. Therapeutic mucosal intranasal vaccination with these peptides in combination with adjuvants significantly reduced HPV tumor growth in mice affording long term survival advantage. Furthermore, combination of intranasal vaccination with the E6 and E7 peptides with immune check point blockade antibody treatmentresulted in significant tumor regression leading to eradication. These data support future clinicaltesting of combination vaccine immunotherapy strategies for effective treatment and possibly eradication of pre- and cancerous HPV lesions.

Michael Underbrink

University of Texas
Slovakia.

Title: Genetic Dysregulation in Recurrent Respiratory Papillomatosis

Time : 18:00-18:20

Biography:

Vladimir Zajac has completed his PhD. in 1982 at the Cancer Research Institute of Slovak Academy of Sciences in Bratislava (Slovakia), where he was from 1996 to 2010 the head of Department of Cancer Genetics. He joined the Medical Faculty of the Comenius University as Associate Professor of Genetics in 2008. He has published 59 papers mostly in reputed journals and he was editor of the book „Bacteria, viruses and parasites in AIDS process“ (InTech, 2011).

Abstract:

Recently accumulate the records, which show that the main site of HIV infection and CD4+ T cell loss is in the GIT and the other mucosal tissue rather than in blood. The HIV-1 was also detected in bowel crypt cells and the lamina propria. It has also been proven that various forms of HIV reservoirs persist in practically all patients receiving HAART. Persistence of HIV was also detected in GALT despite HAART. These facts indicate that in the pathogenesis of AIDS may come into play additional factors. Due to the fact that on mucosa of the colon is a large variety of bacteria and yeast, it is necessary to follow their role in process of pathogenesis. In accordance to this idea, we tested the bacteria and yeast in Slovak and American AIDS patients and HIV-positive children in Kenya and Cambodia for the presence of HIV-like sequences. Using specific primers have been in DNA of these bacteria and yeast identified sequences with high homology to HIV-1. Expression of these sequences has been demonstrated with monoclonal antibodies against p17, p24, p55, gp41 and gp120. Based on these results the presence of HIV-like sequences in bacteria of the patients may be hypothetically explained as follows: 1) bacteria and yeasts are a natural host of HIV sequences; 2) these sequences were transferred into intestinal bacteria from degraded human cells. Hypothetically HIV sequences can be in bacteria and yeast intestinal and respiratory tract present since the beginning of mankind. Thanks to countless epidemics that accompany humanity since its inception, microbes carry these sequences have been largely removed. Administration of antibiotics, drugs and homo and anal sex has recently propagated again. In this way, pathogenic microbes, especially drug resistant, which used to be in the minority moved into the majority. Microbes bearing HIV-like genetic information penetrate from the intestinal tract into the blood. Because of their affinity to lymphocytes, infected or lysed them and a process of immunodeficiency started.

Paola Grandi

University of Pittsburgh
USA.

Title: Highly Selective HSV Virotherapyfor Glioblastoma

Time : 17:00-17:20

Biography:

Dr. Grandi received her Ph.D. from the University of Ferrara (Italy) and was a post-doctoral fellow in the Molecular Neurogenetics Department at the MGH-HMS (Boston). She is now an Assistant Professor in the Dept. of Neurosurgery at the University of Pittsburgh and has a joint appointment in the Department of Microbiology and Molecular Genetics.She has a long standing interest in the molecular biololgy of herpes simplex virus, mechanisms of virus replication and neuropathogenesis and virus host cells interactions that result in innate immune responses to infection

Abstract:

Glioblastoma Multiforme (GBM) is an aggressive brain cancer for which there is no effective treatment. Oncolytic HSV vectors (oHSV) are attenuated lytic viruses that have shown promise in the treatment of human GBM models in animals. Although proven safe for treatment of GBM in patients, oHSVs efficacy has been limited, a consequence of poor intra-tumoral virus replication and spread. To counter these limitations, we have developed oHSVs whose selective replication in GBM cells does not rely on defective genes. This was accomplished by (i) full retargeting of oHSV to utilize the epidermal growth factor receptor (EGFR) for infection of human GBM tumor cells and (ii) further vector engineering to modify the essential HSV immediate early gene (ICP4) for sensitivity to repression by the microRNA mir-124. Mir-124 is highly expressed in neurons but virtually absent in GBM and highly conserved among different species. The mir-124-regulated vector was unable to replicate in nude mice following intracranial inoculation supporting vector safety and was shown to be effective in the treatment of human GBM in nude mice. To enhance vector intra-tumor vector spread, our EGFR retargeted-mir-124 controlled vector was further modified by vector arming with the matrix metalloproteinase gene encoding MMP9. MMP9 degrades collagen type IV, a major component of the extracellular matrix (ECM) and basement membranes of glioblastomas but absent in normal brain tissue. Studies are ongoing to determine whether MMP9 expression enhances vector spread in GBM neurospheres and as a therapeutic agent for enhanced treatment of human GBM in animals.

  • Track 1: General Viral Science
Location: San Antonio, USA

Session Introduction

Charles J. Russell

St. Jude Children’s Research Hospital
USA.

Title: Sendai virus: illuminating parainfluenza virus dynamics in living animals and a platform for vaccine development

Time : 11:25-11:45

Speaker
Biography:

Charles Russell completed his Ph.D. from the University of California at Berkeley where he studied the thermodynamics of membrane-peptide interactions. During his postdoctoral fellowship in the laboratory of Dr. Robert Lamb of the Howard Hughes Medical Institute at Northwestern University, he studied membrane fusion mediated by paramyxoviruses. He is currently a PI at St. Jude Children’s Research Hospital in Memphis, TN, where his lab studies infectious diseases caused by influenza and paramyxoviruses. Research in the Russell lab runs the gamut from high-resolution structure to virus pathogenesis and transmission, making connections between the molecular and biological. Basic research in the lab addresses the impact of fusion glycoprotein activation on pathogenesis and host range while translational work focuses on viral envelope glycoproteins as vaccine candidates.

Abstract:

Sendai virus is the murine counterpart of human parainfluenza virus 1 (HPIV1), the leading cause of pediatric croup. Sendai virus and HPIV1 are antigenically similar, and a team at St. Jude is currently conducting clinical trials to develop Sendai virus as a Jennerian vaccine for HPIV1. Sendai virus is a respiratory paramyxovirus with a single-stranded, negative sense RNA genome that is genetically stable upon inserting a foreign gene (reporter gene or foreign vaccine antigen). In collaboration with our colleagues, we have been using recombinant luciferase-expressing reporter Sendai viruses to visualize the dynamics of infection in living mice and viral glycoprotein-expressing Sendai virus vectors as vaccines for human paramyxoviruses such as RSV and HPIV3. Through the longitudinal measurement of reporter-virus infection as a function of inoculation method or transmission mode (contact or aerosol), our work has revealed that the mode of transmission determines the dynamics of primary respiratory infection and protection from reinfection while independent viral and host factors are responsible for pathogenesis. Basic virological studies on foreign gene positioning and antigen glycoprotein structural form are suggesting novel ways to enhance Sendai virus as a vaccine platform. Our studies with a host-matched virus (Sendai virus in mice) are providing a picture of the elements contributing to natural infection and transmission of a respiratory virus and hint at ways to exploit this understanding to prevent future infections.

Speaker
Biography:

Chuanling Zhang has completed his PhD at the age of 28 years from PekingUnion Medical Collegeand worked at Peking University Health Science Center as an assistant professor. He has published more than 10 papers in reputed journals.

Abstract:

With the aim of studying structure-function relationships of viruses,we developed a general approach that directed azide-bearing amino acids site-specifically to any defined position ofprogeny adeno-associated viruses (AAV)via expanded genetic codes. As a demonstration and application, azides embeddedat AAVcapsids are subsequently coupled withfluorophoresbioorthogonallyin high efficiency, generating visible viruses withprobes incorporated at optimal sites that cause minimal disturbance on viral propagation and infection. The facile and mild realization of site-specific engineering of AAVby natural propagation is of considerable interest to both basic research and for therapeutic applications

Speaker
Biography:

Shirley Komninakis completed her Ph.D. in 2003 and his post-doctorate in 2008. Since then studies the HIV and viral hepatitis. In addition to studying the diversity of viruses currently studying the genome, transcriptome, miRNA and epigenetic of infected patients. Dr. Komninakis is associate professor at the Federal University of São Paulo and Associate Professor at Faculty of Medicine Lusiada Foundation / Santos. These universities have students graduate and undergraduate. In the research laboratory, has expertise in the development of molecular tests involving the Real Time PCR and others.

Abstract:

New antiretroviral agents have been introduced for treatment-experienced patients; one of them is Maraviroc (MVC). MVC is an efficacious entry inhibitor and well tolerated, but it is restricted to patients infected by CCR5 tropic virus. Thus, before using MVC, a tropism test must be performed, as HIV can use the co-receptor CXCR4. The aim of the study was to analyze the genotypic tropism of the variants infecting thirty-six patients failing HAART, receiving multiples regimens and naïve to MVC. Materials and Methods: Thirty-six treatment-experienced patients were enrolled in the study. The average age was 39 years old, the average T CD4+ count was 216 cells/mm³ and the median viral load was 4,63log copies/mL. Nineteen patients live in São Paulo city and seventeen live in Santos city, which has the most important seaport of Latin America. The V3 region of gp120 region was amplified using nested PCR, sequenced and analyzed using the geno2pheno/co-receptor tool (false positive rate cutoff of 10%). Results: Of the 36 patients, 75% (27) were infected with CCR5 tropic variants, while 25% (9) of the patients were infected with CXCR4 variants. Analyzing cities separately, 79% (15) of the patients from São Paulo were infected with the CCR5 tropic virus, while 70% (12) of the patients from Santos city were infected with the CCR5 variants. Conclusions: The genotypic tropism assay is the standard assay for tropism testing and improves the management of treatment in clinical practice. It is known that the use of the CXCR4 co-receptor leads to a rapid disease progression. A high frequency of CCR5 tropic virus were observed among patients on a failing HAART, which indicate that MVC is a good option for treat heavily treatment-experienced patients and that tropism test continue to be a feasible and low cost method to evaluate the tropism of HIV-1 and improve the treatment response

Ming-Liang He

The Chinese University of Hong Kong
China.

Title: Inhibition of enterovirus 71 entry by peptides targeting I β-sheet of VP1 protein

Time : 12:25-12:45

Biography:

Dr He received his Bachelor of Science degree from Sichuan University (Biochemistry). After graduation in 1998, he worked as an engineer in a pharmaceutical factory in Chongqin. He obtained his PhD degree from Shanghai Institute of Biochemistry (and Cell Biology) in 1995. He obtained his postdoctoral training in Roswell Park Cancer Institute at Buffalo (1995–1997) and Washington University School of Medicine at St. Louis in USA (1997–2000). He joined The University of Hong Kong (Institute of Molecular Biology) as Research Assistant Professor in 2000. After SARS outbreak in Hong Kong in 2003, He was appointed as Associate Professor (2005–2011) and Research Associate Professor (2011–2015, Policy Change) at Stanley Ho Center for Emerging Infectious Disease, Faculty of Medicine, The Chinese University of Hong Kong. He joined Department of Biomedical Sciences at City University of Hong Kong in February 2015

Abstract:

The binding of enterovirus capsid protein VP1 on the host receptors is the first step for virus entry during the infection. Peptides blocking the binding of VP1 and its receptors can be used not only for antiviral treatment but also for probing functional important motifs or residues involved in this biological process. Based on the crystal structure of VP1-VP4 complex of enterovirus 71 (EV71), we applied molecular modeling approaches to revisit the potential antiviral peptides in an anti-enterovirus 71 screening study. We showed that a β-sheet structure (I β) is unique and locates at a very favorable position for drug target. More importantly, the sequence of I β-sheet is highly conserved among subtypes of EV71, suggesting an idea target sites for antiviral drug design. Peptides targeting I β-sheet potently inhibited EV71 infections. Further attachment and single-round pseudovirus infection assays revealed that the attachment of virions on host cells was effectively blocked by peptides. Alanine scan analysis demonstrated that residues Arg250, Arg254, Met255 and Lys256 are critical for virion binding on host cells. This study demonstrated the importance of I β-sheet structure for EV71 entry and an effective peptide for block virion-host cell interactions.